Modified dog alpha1-interferon gene, construction method of expression vector and preparation method of protein encoded by modified dog alpha1-interferon gene
An expression vector and interferon technology, applied in the field of construction and protein preparation, can solve problems such as unfavorable industrial production of canine α1-interferon, no technical solution, and inability to fundamentally solve the cost problem of canine α1-interferon , to achieve the effect of improved antiviral activity, low cost and simple method
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[0051] 1. Design and synthesize a pair of primers according to the nucleotide sequence of canine α1-interferon to synthesize canine α1-interferon gene;
[0052] The present invention refers to the literature (Gurkan C, 2005; Zhao Xiang, 2000) to analyze the codon preference of the Pichia pastoris expression system, and bias the whole gene sequence of canine interferon α1 published in Genbank (A33693) Transformation of addiction, while adding separately upstream and downstream EcoR I and Xba I restriction site (underlined part) and protective base, and in EcoR A base G is added upstream of the I restriction site. The modified gene was synthesized by Shanghai Bioengineering Co., Ltd., and the modified canine α1-interferon gene was inserted into the multiple cloning site of plasmid pUC57. The recombinant plasmid was named pUC57-CaIFN-α1. According to the modified canine interferon α1 nucleotide sequence, a pair of primers are designed and synthesized:
[0053] ...
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